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Immunogold Electron Microscopy

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.

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Detached Maize Sheaths for Live-Cell Imaging of Infection by Fungal Foliar Maize Pathogens

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Published on: September 15, 2023

Protein immunolocalization in maize tissues.

Cristian Forestan¹, Nicola Carraro, Serena Varotto

¹Department of Agronomy, Food, Natural Resources, Animals and Environment, University of Padova, Padova, Italy.

Methods in Molecular Biology (Clifton, N.J.)

|January 10, 2013

Summary

This study details a robust immunolocalization protocol for plant protein analysis in maize. It enables precise cellular and subcellular protein localization, crucial for understanding plant development.

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Area of Science:

Plant developmental biology
Molecular biology
Cell biology

Background:

Analyzing gene expression at transcript and protein levels is vital for plant development.
Protein localization studies use immunolocalization with antibodies or fluorescent protein fusions.
Immunolocalization confirms reporter gene expression and offers an alternative to fusion proteins, especially in maize.

Purpose of the Study:

To describe a complete procedure for protein localization in maize tissues at cellular and subcellular levels.
To provide a method for studying protein targeting to different cellular compartments.
To present a protocol for analyzing nuclear histone distribution in maize root apexes.

Main Methods:

Utilizing polyclonal or monoclonal antibodies for immunolocalization.
Embedding plant tissues in paraplast, PEG400, or agarose for microscopy.
Employing epifluorescence and confocal microscopy for observations.
Developing a protocol for squashed maize root apexes to analyze nuclear histone distribution.

Main Results:

Established a comprehensive immunolocalization protocol for diverse maize tissues.
Demonstrated the effectiveness of immunolocalization for subcellular protein targeting studies.
Successfully analyzed nuclear histone distribution in maize root apexes.

Conclusions:

Immunolocalization is a powerful tool for studying protein localization in maize.
The described methods facilitate detailed analysis of protein targeting and nuclear distribution.
This protocol is valuable for plant developmental biology research, particularly in maize.